Nerve Guidance Conduits Research Articles

Does Electrical Stimulation through Nerve Conduits Improve Peripheral Nerve Regeneration?-A Systematic Review.

Peripheral nerve injuries affect over 2% of trauma patients and can lead to severe functional impairment and permanent disability. Autologous nerve transplantation is still the gold standard in the reconstruction of nerve defects. For small defects, conduits can be considered for bridging. Lately, the combined use of conduits and electrical stimulation has gained attention in the treatment of peripheral nerve injury. This review aimed to present the currently available data on this topic. PubMed, Embase, Medline and the Cochrane Library were searched for studies on electrical stimulation through nerve conduits for nerve defects in in vivo studies. Fifteen studies fit the inclusion criteria. All of them reported on the application of nerve conduits combined with stimulation for sciatic nerve gaps in rats. Functional, electrophysiological and histological evaluations showed improved nerve regeneration after electrical stimulation. High variation was observed in the treatment protocols. Electrically stimulated conduits could improve peripheral nerve regeneration in rat models. The combined application of nerve guidance conduits and electrical stimulation shows promising results and should be further evaluated under standardized conditions.

Silk-in-Silk Nerve Guidance Conduits Enhance Regeneration in a Rat Sciatic Nerve Injury Model.

Advanced nerve guidance conduits can provide an off-the-shelf alternative to autografts for the rehabilitation of segmental peripheral nerve injuries. In this study, the excellent processing ability of silk fibroin and the outstanding cell adhesion quality of spider dragline silk are combined to generate a silk-in-silk conduit for nerve repair. Fibroin-based silk conduits (SC) are characterized, and Schwann cells are seeded on the conduits and spider silk. Rat sciatic nerve (10mm) defects are treated with an autograft (A), an empty SC, or a SC filled with longitudinally aligned spider silk fibers (SSC) for 14weeks. Functional recovery, axonal re-growth, and re-myelination are assessed. The material characterizations determine a porous nature of the conduit. Schwann cells accept the conduit and spider silk as growth substrate. The in vivo results show a significantly faster functional regeneration of the A and SSC group compared to the SC group. In line with the functional results, the histomorphometrical analysis determines a comparable axon density of the A and SSC groups, which is significantly higher than the SC group. These findings demonstrate that the here introduced silk-in-silk nerve conduit achieves a similar regenerative performance as autografts largely due to the favorable guiding properties of spider dragline silk.

3D Printed Conductive Multiscale Nerve Guidance Conduit with Hierarchical Fibers for Peripheral Nerve Regeneration.

Nerve guidance conduits (NGCs) have become a promising alternative for peripheral nerve regeneration; however, the outcome of nerve regeneration and functional recovery is greatly affected by the physical, chemical, and electrical properties of NGCs. In this study, a conductive multiscale filled NGC (MF-NGC) consisting of electrospun poly(lactide-co-caprolactone) (PCL)/collagen nanofibers as the sheath, reduced graphene oxide /PCL microfibers as the backbone, and PCL microfibers as the internal structure for peripheral nerve regeneration is developed. The printed MF-NGCs presented good permeability, mechanical stability, and electrical conductivity, which further promoted the elongation and growth of Schwann cells and neurite outgrowth of PC12 neuronal cells. Animal studies using a rat sciatic nerve injury model reveal that the MF-NGCs promote neovascularization and M2 transition through the rapid recruitment of vascular cells and macrophages. Histological and functional assessments of the regenerated nerves confirm that the conductive MF-NGCs significantly enhance peripheral nerve regeneration, as indicated by improved axon myelination, muscle weight increase, and sciatic nerve function index. This study demonstrates the feasibility of using 3D-printed conductive MF-NGCs with hierarchically oriented fibers as functional conduits that can significantly enhance peripheral nerve regeneration.

Topography and mechanical measurements of primary Schwann cells and neuronal cells with atomic force microscope for understanding and controlling nerve growth

Peripheral nerve injuries require a piece of substantial information for a satisfactory treatment. The prior peripheral nerve injury knowledge, can improve nerve repair, and its growth at molecular and cellular level. In this study, we employed an atomic force microscope (AFM) to investigate the topography and mechanical properties of the primary Schwann cells and neuronal cells. Tapping mode images and contact points force-volume maps provide the cells topography. Two different probes were used to acquire the micro and nanomechanical properties of the primary Schwann cells, NG-108–15 neuronal cells, and growth cones. Moreover, the sharp probe was only used to investigate neurites nanomechanics. A significant difference in the elastic moduli found between primary Schwann cells, and neuronal cells, with both probes, with consistent results. The elastic moduli of the growth cones were found higher, than the neuronal cells and primary Schwann cells, with both probes. Furthermore, the modulus variations were also found between neurites. These results have significant implications for a better understanding of the peripheral nerve system (PNS) in terms of defining the optimal pattern surface and nerve guidance conduits.

Correction to: Heparin/Growth Factors-Immobilized Aligned Electrospun Nanofibers Promote Nerve Regeneration in Polycaprolactone/Gelatin-Based Nerve Guidance Conduits

Correction to: Heparin/Growth Factors-Immobilized Aligned Electrospun Nanofibers Promote Nerve Regeneration in Polycaprolactone/Gelatin-Based Nerve Guidance Conduits

Additive Manufacturing of Polyhydroxyalkanoate-Based Blends Using Fused Deposition Modelling for the Development of Biomedical Devices.

In the last few decades Additive Manufacturing has advanced and is becoming important for biomedical applications. In this study we look at a variety of biomedical devices including, bone implants, tooth implants, osteochondral tissue repair patches, general tissue repair patches, nerve guidance conduits (NGCs) and coronary artery stents to which fused deposition modelling (FDM) can be applied. We have proposed CAD designs for these devices and employed a cost-effective 3D printer to fabricate proof-of-concept prototypes. We highlight issues with current CAD design and slicing and suggest optimisations of more complex designs targeted towards biomedical applications. We demonstrate the ability to print patient specific implants from real CT scans and reconstruct missing structures by means of mirroring and mesh mixing. A blend of Polyhydroxyalkanoates (PHAs), a family of biocompatible and bioresorbable natural polymers and Poly(L-lactic acid) (PLLA), a known bioresorbable medical polymer is used. Our characterisation of the PLA/PHA filament suggest that its tensile properties might be useful to applications such as stents, NGCs, and bone scaffolds. In addition to this, the proof-of-concept work for other applications shows that FDM is very useful for a large variety of other soft tissue applications, however other more elastomeric MCL-PHAs need to be used.

Additive manufacturing of Schwann cell-laden collagen/alginate nerve guidance conduits by freeform reversible embedding regulate neurogenesis via exosomes secretion towards peripheral nerve regeneration

Peripheral nerve injury is a common clinical problem that could be debilitating to one's quality of life. The complex nerve guidance conduits (NGCs) with cells in order to improve nerve regeneration. Therefore, we used freeform reversible embedding of suspended hydrogels to fabricate Schwann cells (SCs)-laden collagen/alginate (Col/Alg) NGCs. First, we evaluated Col influence on the characteristics of NGCs. After which, Wharton's jelly mesenchymal stem cells (WJMSC) are seeded onto the inner channel of NGCs and evaluated neural regeneration behaviors. Results indicated the SCs-laden NGCs with 2.5 % Col found the highest proliferation and secretion of neurotrophic protein. Furthermore, co-culture of SCs promoted differentiation of WJMSC as seen from the increased neurogenic-related protein in NGCs. To determine the molecular mechanism between SCs and WJMSC, we demonstrated the neurotrophic factors secreted by SCs act on tropomyosin receptor kinase A (TrkA) receptors of WJMSC to promote nerve regeneration. In addition, our study demonstrated SCs-derived exosomes had a critical role in regulating neural differentiation of WJMSC. Taken together, this study demonstrates the fabrication of SCs-laden Col/Alg NGCs for nerve regeneration and understanding regarding the synergistic regenerative mechanisms of different cells could bring us a step closer for clinical treatment of large nerve defects.

Artificial nerve graft constructed by coculture of activated Schwann cells and human hair keratin for repair of peripheral nerve defects.

Studies have shown that human hair keratin (HHK) has no antigenicity and excellent mechanical properties. Schwann cells, as unique glial cells in the peripheral nervous system, can be induced by interleukin-1β to secrete nerve growth factor, which promotes neural regeneration. Therefore, HHK with Schwann cells may be a more effective approach to repair nerve defects than HHK without Schwann cells. In this study, we established an artificial nerve graft by loading an HHK skeleton with activated Schwann cells. We found that the longitudinal HHK microfilament structure provided adhesion medium, space and direction for Schwann cells, and promoted Schwann cell growth and nerve fiber regeneration. In addition, interleukin-1β not only activates Schwann cells, but also strengthens their activity and increases the expression of nerve growth factors. Activated Schwann cells activate macrophages, and activated macrophages secrete interleukin-1β, which maintains the activity of Schwann cells. Thus, a beneficial cycle forms and promotes nerve repair. Furthermore, our studies have found that the newly constructed artificial nerve graft promotes the improvements in nerve conduction function and motor function in rats with sciatic nerve injury, and increases the expression of nerve injury repair factors fibroblast growth factor 2 and human transforming growth factor B receptor 2. These findings suggest that this artificial nerve graft effectively repairs peripheral nerve injury.

Engineered multifunctional silk fibroin/gelatin hydrogel conduit loaded with miR-29a@ZIF-8 nanoparticles for peripheral nerve regeneration

Peripheral nerve injury (PNI) is a common surgical disease. In recent years, with the development of tissue engineering materials, nerve guidance conduit (NGC) is expected to replace autologous nerve transplantation and become a new method for the treatment of PNI. In this work, we developed a multifunctional silk fibroin (SF)/gelatin-tyramine (GT) composite hydrogel conduit with flexible adjustable size by using a diffusion-driven cross-linking method. Furthermore, the ZIF-8 nanoparticles loaded with miR-29a (miR-29a@ZIF-8) delivery system was constructed and compounded into SF/GT hydrogel conduit to enhance its bioactivity and neural repair effects through sustained miR-29a release. In vitro cell experiments showed that SF/GT hydrogel conduit could significantly promote the myelination of Schwann cells (SCs), neuronal differentiation and axon extension of PC12 ​cells. In addition, it was worth mentioning that SF/GT hydrogel conduit could also regulate the immune microenvironment of nerve regeneration by promoting the transformation of macrophages from M1 phenotype to M2 phenotype, indicating a potential application as nerve guidance conduit in peripheral nerve repair.

A hyaluronic acid granular hydrogel nerve guidance conduit promotes regeneration and functional recovery of injured sciatic nerves in rats.

A hyaluronic acid granular hydrogel can promote neuronal and astrocyte colony formation and axonal extension in vitro, suggesting that the hydrogel can simulate an extracellular matrix structure to promote neural regeneration. However, in vivo experiments have not been conducted. In this study, we transplanted a hyaluronic acid granular hydrogel nerve guidance conduit to repair a 10-mm long sciatic nerve gap. The Basso, Beattie, and Bresnahan locomotor rating scale, sciatic nerve compound muscle action potential recording, Fluoro-Gold retrograde tracing, growth related protein 43/S100 immunofluorescence staining, transmission electron microscopy, gastrocnemius muscle dry/wet weight ratio, and Masson's trichrome staining results showed that the nerve guidance conduit exhibited similar regeneration of sciatic nerve axons and myelin sheath, and recovery of the electrophysiological function and motor function as autologous nerve transplantation. The conduit results were superior to those of a bulk hydrogel or silicone tube transplant. These findings suggest that tissue-engineered nerve conduits containing hyaluronic acid granular hydrogels effectively promote the morphological and functional recovery of the injured sciatic nerve. The nerve conduits have the potential as a material for repairing peripheral nerve defects.

Biodegradable polyurethane-incorporating decellularized spinal cord matrix scaffolds enhance Schwann cell reprogramming to promote peripheral nerve repair.

Decellularized extracellular matrix (dECM) nerve guide conduits (NGCs) are a promising strategy to replace autogenous nerve grafting for the treatment of peripheral nerve system (PNS) injury. However, dECM conduits with mechanical properties that match those of peripheral nerves are yet to be well developed. Herein, we developed polyurethane-based NGCs incorporating decellularized spinal cord (BWPU-DSC NGCs) to repair peripheral nerves. BWPU-DSC NGCs have an inner three-dimensional micro-nanostructure. The mechanical properties of BWPU-DSC NGCs were similar to those of polyurethane NGCs, which were proven to promote peripheral nerve regeneration. An in vitro study indicated that BWPU-DSC NGCs could boost the proliferation and growth of cell processes in Schwann and neuron-like cells. In a rat sciatic nerve transected injury model, BWPU-DSC NGCs exhibited a dramatic increase in nerve repair, similar to that obtained by the current gold standard autograft implantation at only 6 weeks post-implantation, whereas polyurethane NGCs still displayed incomplete nerve repair. Histological analysis revealed that BWPU-DSC NGCs could induce the reprogramming of Schwann cells to promote axon regeneration and remyelination. Moreover, reprogrammed Schwann cells together with BWPU-DSC NGCs had anti-inflammatory effects and altered the activation state of macrophages to M2 phenotypes to enhance PNS regeneration. In this study, we provided a strategy to prepare polyurethane-based dECM NGCs enriched with bioactive molecules to promote PNS regeneration.

Phosphorene hydrogel conduits as "neurotrophin reservoirs" for promoting regeneration of peripheral nerves.

Treatment of large gaps in peripheral nerves is a major clinical challenge. Artificial nerve guidance conduits (NGCs) have provided new opportunities for guiding nerve regeneration. In this study, multifunctional black phosphorus (BP) hydrogel NGCs loaded with neuregulin 1 (Nrg1) were fabricated to support peripheral-nerve regeneration: they exhibited good flexibility and nerve regeneration-related cell induction, promoted Schwann-cell proliferation and accelerated neuron-branch elongation. Nrg1 induced the proliferation and migration of Schwann cells, which had beneficial roles in promoting nerve regeneration. In vivo immunofluorescence studies revealed BP hydrogel NGCs loaded with Nrg1 promoted sciatic-nerve regeneration and axon remyelination. Our method has great potential for promoting treatment of peripheral-nerve injuries.

The best of both worlds: mastering nerve regeneration combining biological and nanotechnological tools.

The best of both worlds: mastering nerve regeneration combining biological and nanotechnological tools.

Nerve Regeneration Effect of a Composite Bioactive Carboxymethyl Chitosan-Based Nerve Conduit with a Radial Texture.

Chitosan (CTS) has been used as a nerve guidance conduit (NGC) material for bridging peripheral nerve defects due to its biocompatible, biodegradable, and non-toxic properties. However, the nerve regeneration effect of chitosan alone is restricted due to its inadequate biological activity. Herein, a composite, bioactive chitosan based nerve conduit, consisting of outer warp-knitted tube scaffold made from medical-grade chitosan fiber, and inner porous cross linked carboxymethyl chitosan (C-CM-CTS) sponge with radial texture was developed. The inner wall of the scaffold was coated with C-CM-CTS solution. CM-CTS provided favorable bioactivities in the composite chitosan-based nerve conduit. An in vitro study of CM-CTS revealed its satisfying biocompatibility with fibroblast and its inhibition of oxidative damage to Schwann cells. As the internal filler of the NGC, the lyophilized sponge of C-CM-CTS showed a longitudinal guidance effect for nerve reconstruction. After 10 mm defect in rat sciatic nerve was bridged with the composite bioactive chitosan-based nerve conduit, the nerve conduit was able to effectively promote axonal regeneration and played a positive role in inducing nerve regeneration and functional recovery. In addition to the functional advantages, which are equal to those of an autograft; the technology for the preparation of this conduit can be put into mass production.

3D-bioprinted GelMA nerve guidance conduits promoted peripheral nerve regeneration by inducing trans-differentiation of MSCs into SCLCs via PIEZO1/YAP axis

Schwann cells (SCs)-based nerve guidance conduits (NGCs) is a promising strategy for repairing long-gap peripheral nerve injury (PNI). But the number of SCs is limited as terminally differentiated cells. Matrix stiffness is able to direct cell fate of stem cells. And nerve-special stiffness (NSS) may contribute to stem cells converting into SCs. However, the potential mechanisms remain uncertain. The present study aimed to elucidate whether the given NSS was able to promote mesenchymal stem cells (MSCs) trans-differentiating into Schwann cell-like cells (SCLCs), facilitating cell-based NGCs repairing PNI. Gelatin methacryloylate (GelMA) hydrogels with different stiffness were manufactured, and their roles in the trans-differentiation of MSCs into SCLCs were investigated in vitro study. The most favorable stiffness (0.9–2.9 kPa) for trans-differentiation and neurotrophic factors expression was confirmed by western blot and immunofluorescence assays. The reason might lie in softer stiffness stretched cell morphology, leading to up-regulation of PIEZO1 and then activating YAP nuclear translocation. As for in vivo study, multi-channel NGCs containing SCLCs were fabricated with extrusion-based bioprinting and then filled a 5 mm gap in sciatic nerve defect of SD rats. It turned out that GelMA-made NGCs with stiffness of 2.9 kPa achieved promising neurogenerative capacity, not only promoting recovering sensory and motor functions, but also improving myelinated nerve fiber regeneration. Our findings demonstrated that 0.9–2.9 kPa might be the desired mechanical strength inducing trans-differentiation into SCLCs via PIEZO1/YAP axis, and the 3D-printed GelMA NGCs combined with SCLCs could be a potential candidate for long-gap peripheral nerve injury.

Combining a Density Gradient of Biomacromolecular Nanoparticles with Biological Effectors in an Electrospun Fiber-Based Nerve Guidance Conduit to Promote Peripheral Nerve Repair.

Peripheral nerve injury is a serious medical problem with limited surgical and clinical treatment options. It is of great significance to integrate multiple guidance cues in one platform of nerve guidance conduits (NGCs) to promote axonal elongation and functional recovery. Here, a multi-functional NGC is constructed to promote nerve regeneration by combining ordered topological structure, density gradient of biomacromolecular nanoparticles, and controlled delivery of biological effectors to provide the topographical, haptotactic, and biological cues, respectively. On the surface of aligned polycaprolactone nanofibers, a density gradient of bioactive nanoparticles capable of delivering recombinant human acidic fibroblast growth factor is deposited. On the graded scaffold, the proliferation of Schwann cells is promoted, and the directional extension of neurites from both PC12 cells and dorsal root ganglions is improved in the direction of increasing particle density. After being implanted in vivo for 6 and 12 weeks to repair a 10-mm rat sciatic nerve defect, the NGC promotes axonal elongation and remyelination, achieving the regeneration of the nerve not only in anatomical structure but also in functional recovery. Taken together, the NGC provides a favorable microenvironment for peripheral nerve regeneration and holds great promise for realizing nerve repair with an efficacy close to autograft.

EMSCs‐Seeded Micro‐Stripe Patterned Polycaprolactone Promoting Sciatic Nerve Regeneration

AbstractTransplantation of a synthetic polymer nerve conduit carrying stem cells is promising for treating peripheral nerve injury. In this study, a micropatterned inner surface of polycaprolactone (PCL) nerve conduit cellularized with nasal respiratory mucosa‐derived ectoderm mesenchymal stem cells (EMSCs) is adopted for peripheral nerve regeneration. The PCL microstructure grating is fabricated by soft nanoimprint lithography (NIL), whose templates can be easily transferred from the Ebeam photoresist pattern. The most suitable microstructure is carefully chosen to support EMSCs alignment and nerve regeneration. In vitro, the micropatterned PCL nerve conduit greatly enhanced the directional alignment of EMSCs and the MBP and S100 expression of myelin proteins. Additionally, the micropatterned conduit preseeded with EMSCs significantly stimulated the axon‐directed extension of neural stem cells. In vivo, micropatterned PCL conduits cotransplanted with EMSCs enhanced motor function and promoted sciatic nerve regeneration and extension in a rat model of sciatic nerve defect. Taken together, this “all‐in‐one” nerve guidance conduit makes sciatic nerve regeneration and morphological repairing accelerated, which has a potential clinical application in neural tissue engineering.

Neuronal cell response on aligned fibroporous electrospun mat generated from silver ion complexed ethylene vinyl alcohol copolymer.

Generating electrospun mats with aligned fibers and obtaining neurite extension in the aligned fiber direction could provide hope for fabricating nerve guidance conduits or wraps through an easy method. The growing interest in generating electrospun mats with aligned fibers for tissue engineering is looking for simple methods to generate the same. Here, in this study, ethylene vinyl alcohol copolymer (EVAL) chains were complexed with silver ions (Ag+ ) to generate aligned fibers during the electrospinning process. The fibers thus produced were subjected to physico-chemical characterization and biological studies to ensure their properties and to examine whether suitable for neuronal cell attachment and neurite extension that may be useful in making nerve guidance conduits or wraps. The presence of silver ions and its complex formation with -OH of EVAL has been confirmed with EDX and XPS analysis respectively. The alignment of fibers was visualized from SEM analysis and confirmed using directionality analysis using Fiji-ImageJ software. Mechanical properties done with dumbbells punched out in longitudinal and transverse directions also substantiated the alignment of fibers. The results obtained from direct contact, MTT, and live/dead assay showed the cells are viable on the material. From the actin staining and immunostaining assays, it was evident that the PC12 cells could attach and extend their neurites in an aligned manner on the fibers. The maximum neurite extension was up to 200 μm in length. These properties of electrospun EVAL-Ag mat with aligned fibers indicated that it could be developed as a biocompatible nerve guidance conduit or wrap.

Multifaceted polymeric nerve guidance conduits with distinctive double-layered architecture and plasma-induced inner chemistry gradient for the repair of critical-sized defects

Despite tissue engineering advances, current nerve guidance conduits (NGCs) are still failing in repairing critical-sized defects. This study aims, therefore, at tackling large nerve gaps (2 cm) by designing NGCs possessing refined physicochemical properties enhancing the activity of Schwann cells (SCs) that support nerve regeneration over long distances. As such, a combinatorial strategy adopting novel plasma-induced surface chemistry and architectural heterogeneity was considered. A mechanically suitable copolymer (Polyactive®) was electrospun to produce nanofibrous NGCs mimicking the extracellular matrix. An innovative seamless double-layered architecture consisting of an inner wall comprised of bundles of aligned fibers with intercalated random fibers and an outer wall fully composed of random fibers was conceived to synergistically provide cell guidance cues and sufficient nutrient inflow. NGCs were subjected to argon plasma treatments using a dielectric barrier discharge (DBD) and a plasma jet (PJ). Surface chemical changes were examined by advanced X-ray photoelectron spectroscopy (XPS) micro-mappings. The DBD homogeneously increased the surface oxygen content from 17 % to 28 % on the inner wall. The PJ created a gradient chemistry throughout the inner wall with an oxygen content gradually increasing from 21 % to 30 %. In vitro studies revealed enhanced primary SC adhesion, elongation and proliferation on plasma-treated NGCs. A cell gradient was observed on the PJ-treated NGCs thus underlining the favorable oxygen gradient in promoting cell chemotaxis. A gradual change from circular to highly elongated SC morphologies mimicking the bands of Büngner was visualized along the gradient. Overall, plasma-treated NGCs are promising candidates paving the way towards critical nerve gap repair.

Construction of a mineralized collagen nerve conduit for peripheral nerve injury repair.

A new nerve guidance conduits (NGCs) named MC@Col containing Type I collagen (Col) and mineralized collagen (MC) was developed, enhancing mechanical and degradation behavior. The physicochemical properties, the mechanical properties and in vitro degradation behavior were all evaluated. The adhesion and proliferation of Schwann cells (SCs) were observed. In the in vivo experiment, MC@Col NGC and other conduits including Col, chitosan (CST) and polycaprolactone (PCL) conduit were implanted to repair a 10-mm-long Sprague-Dawley rat's sciatic nerve defect. Histological analyses, morphological analyses, electrophysiological analyses and further gait analyses were all evaluated after implantation in 12 weeks. The strength and degradation performance of the MC@Col NGC were improved by the addition of MC in comparison with pure Col NGC. In vitro cytocompatibility evaluation revealed that the SCs had good viability, attachment and proliferation in the MC@Col. In in vivo results, the regenerative outcomes of MC@Col NGC were close to those by an autologous nerve graft in some respects, but superior to those by Col, CST and PCL conduits. The MC@Col NGC exhibited good mechanical performance as well as biocompatibility to bridge nerve gap and guide nerve regeneration, thus showing great promising potential as a new type of conduit in clinical applications.